The Astrocyte Nexus: CX 43-Protein Interactions

描述(申请人提供)：远距离细胞间通信 星形胶质细胞之间很大程度上是由缝隙连接通道通过 这些离子和代谢物直接从一个细胞传递到下一个细胞。黄瓜的毛孔 星形胶质细胞缝隙连接主要由缝隙连接蛋白组成 连接蛋白43。最近的证据表明，其他蛋白质与 Cx43在缝隙连接处，我们将这种大分子复合体命名为 Nexus。我们假设Nexus组件可能调节两个 缝隙连接通道的特性，也可能在细胞内发挥作用 信号转导。为了检验这一假设，我们将确定 星形胶质细胞中与Cx43结合的其他蛋白质的鉴定 转基因细胞，实现高通量和定量的方法 识别新的Nexus蛋白质，测量相互作用的强度，以及 使用Cx43突变体和Cx43结合进行生理实验 合作伙伴，以确定这种互动的功能后果。在……里面 此外，此应用程序的一个主要目标是获取结构信息 关于Cx43相互作用和相互作用的结构域 蛋白质。这些研究使用了多学科的方法，旨在探索 这一领域的新概念有望带来小说 了解缝隙连接在神经系统和 其他地方。

DESCRIPTION (provided by the applicant): Long range intercellular communication among astrocytes is in large part mediated by gap junction channels through which ions and metabolites pass directly from one cell to the next. The pore of astrocyte gap junctions is formed primarily of the gap junction protein connexin43. Recent evidence indicates that other proteins are associated with Cx43 at gap junctions, and we have termed this macromolecular complex the Nexus. We hypothesize that the Nexus components may regulate both the properties of the gap junction channels and also may function in intracellular signal transduction. In order to test this hypothesis, we will determine the identities of the other proteins that bind to Cx43 in astrocytes and in transfected cells, implement high throughput and quantitative methods to identify novel Nexus proteins, measure the strengths of interaction, and perform physiological experiments with Cx43 mutants and with Cx43 binding partners to determine the functional consequences of such interactions. In addition, a major goal of this application is to obtain structural information regarding domains of Cx43 that interact with each other and with other proteins. These studies use a multidisciplinary approach directed at exploring a new concept in the field and as such are expected to lead to novel understanding of roles that gap junctions play in the nervous system and elsewhere.